PMID- 38048933
OWN - NLM
STAT- MEDLINE
DCOM- 20240129
LR  - 20240206
IS  - 1879-0003 (Electronic)
IS  - 0141-8130 (Linking)
VI  - 257
IP  - Pt 2
DP  - 2024 Feb
TI  - Biochemical characterization of glutaminase-free L-asparaginases from Himalayan 
      Pseudomonas and Rahnella spp. for acrylamide mitigation.
PG  - 128576
LID - S0141-8130(23)05475-2 [pii]
LID - 10.1016/j.ijbiomac.2023.128576 [doi]
AB  - L-asparaginase having low glutaminase activity is important in clinical and food 
      applications. Herein, glutaminase-free L-asparaginase (type I) coding genes from 
      Pseudomonas sp. PCH182 (Ps-ASNase I) and Rahnella sp. PCH162 (Rs-ASNase I) was 
      amplified using gene-specific primers, cloned into a pET-47b(+) vector, and 
      plasmids were transformed into Escherichia coli (E. coli). Further, affinity 
      chromatography purified recombinant proteins to homogeneity with monomer sizes of 
      ~37.0 kDa. Purified Ps-ASNase I and Rs-ASNase I were active at wide pHs and 
      temperatures with optimum activity at 50  degrees C (492 +/- 5 U/mg) and 37  degrees C 
      (308 +/- 4 U/mg), respectively. Kinetic constant K(m) and V(max) for L-asparagine 
      (Asn) were 2.7 +/- 0.06 mM and 526.31 +/- 4.0 U/mg for Ps-ASNase I, and 
      4.43 +/- 1.06 mM and 434.78 +/- 4.0 U/mg for Rs-ASNase I. Circular dichroism study 
      revealed 29.3 % and 24.12 % alpha-helix structures in Ps-ASNase I and Rs-ASNase I, 
      respectively. Upon their evaluation to mitigate acrylamide formation, 43 % and 
      34 % acrylamide (AA) reduction were achieved after pre-treatment of raw potato 
      slices, consistent with 65 % and 59 % Asn reduction for Ps-ASNase I and Rs-ASNase 
      I, respectively. Current findings suggested the potential of less explored 
      intracellular L-asparaginase in AA mitigation for food safety.
CI  - Copyright (c) 2023 Elsevier B.V. All rights reserved.
FAU - Patial, Vijeta
AU  - Patial V
AD  - Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, 
      Palampur 176 061, Himachal Pradesh, India; Academy of Scientific & Innovative 
      Research (AcSIR), Ghaziabad 201 002, India.
FAU - Kumar, Subhash
AU  - Kumar S
AD  - Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, 
      Palampur 176 061, Himachal Pradesh, India; Academy of Scientific & Innovative 
      Research (AcSIR), Ghaziabad 201 002, India.
FAU - Joshi, Robin
AU  - Joshi R
AD  - Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, 
      Palampur 176 061, Himachal Pradesh, India.
FAU - Singh, Dharam
AU  - Singh D
AD  - Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, 
      Palampur 176 061, Himachal Pradesh, India; Academy of Scientific & Innovative 
      Research (AcSIR), Ghaziabad 201 002, India. Electronic address: 
      dharamsingh@ihbt.res.in.
LA  - eng
PT  - Journal Article
DEP - 20231202
PL  - Netherlands
TA  - Int J Biol Macromol
JT  - International journal of biological macromolecules
JID - 7909578
RN  - EC 3.5.1.1 (Asparaginase)
RN  - EC 3.5.1.2 (Glutaminase)
RN  - 20R035KLCI (Acrylamide)
RN  - 7006-34-0 (Asparagine)
RN  - 0 (Antineoplastic Agents)
SB  - IM
MH  - Asparaginase/chemistry
MH  - *Rahnella/metabolism
MH  - Escherichia coli/genetics/metabolism
MH  - Pseudomonas/genetics/metabolism
MH  - Glutaminase/genetics
MH  - Acrylamide
MH  - Asparagine/metabolism
MH  - *Antineoplastic Agents
OTO - NOTNLM
OT  - Acrylamide
OT  - Enzyme kinetics
OT  - Heterologous expression
OT  - L-asparaginase
OT  - Potato chips
OT  - Thermostability
COIS- Declaration of competing interest The authors declare no conflict of personal or 
      financial interest.
EDAT- 2023/12/05 00:41
MHDA- 2024/01/29 06:43
CRDT- 2023/12/04 19:24
PHST- 2023/07/07 00:00 [received]
PHST- 2023/11/20 00:00 [revised]
PHST- 2023/12/01 00:00 [accepted]
PHST- 2024/01/29 06:43 [medline]
PHST- 2023/12/05 00:41 [pubmed]
PHST- 2023/12/04 19:24 [entrez]
AID - S0141-8130(23)05475-2 [pii]
AID - 10.1016/j.ijbiomac.2023.128576 [doi]
PST - ppublish
SO  - Int J Biol Macromol. 2024 Feb;257(Pt 2):128576. doi: 
      10.1016/j.ijbiomac.2023.128576. Epub 2023 Dec 2.